Biotin synthase contains two distinct iron-sulfur cluster binding sites: chemical and spectroelectrochemical analysis of iron-sulfur cluster interconversions.
AuthorsUgulava NB, Gibney BR, Jarrett JT
JournalBiochemistry
PubMed ID11444981
Biotin synthase is an iron-sulfur protein that utilizes AdoMet to catalyze the presumed radical-mediated insertion of a sulfur atom between the saturated C6 and C9 carbons of dethiobiotin. Biotin synthase (BioB) is aerobically purified as a dimer that contains [2Fe-2S](2+) clusters and is inactive in the absence of additional iron ... More
Easily reversible desthiobiotin binding to streptavidin, avidin, and other biotin-binding proteins: uses for protein labeling, detection, and isolation.
AuthorsHirsch JD, Eslamizar L, Filanoski BJ, Malekzadeh N, Haugland RP, Beechem JM, Haugland RP
JournalAnal Biochem
PubMed ID12419349
'The high-affinity binding of biotin to avidin, streptavidin, and related proteins has been exploited for decades. However, a disadvantage of the biotin/biotin-binding protein interaction is that it is essentially irreversible under physiological conditions. Desthiobiotin is a biotin analogue that binds less tightly to biotin-binding proteins and is easily displaced by ... More
Diaminobiotin and desthiobiotin have biotin-like activities in Jurkat cells.
AuthorsRodriguez-Melendez R, Lewis B, McMahon RJ, Zempleni J
JournalJ Nutr
PubMed ID12730407
'In mammals, biotin serves as a coenzyme for carboxylases such as propionyl-CoA carboxylase. The expression of genes encoding interleukin-2 (IL-2) and IL-2 receptor (IL-2R)gamma also depends on biotin. Biotin metabolites are structurally similar to biotin, and their concentrations in tissues are quantitatively important. Here, the hypothesis was tested that biotin ... More
Anterograde tracing of retinal axons in the avian embryo with low molecular weight derivatives of biotin.
AuthorsHalfter W
JournalDev Biol
PubMed ID2433173
'Several reactive biotin esters were injected into the eyes of chick and quail embryos at various stages of development. Four of the biotin esters reacted with molecules of the eye tissue and were detected with light and electron microscopy in fluorescein isothiocyanate and peroxidase-avidin incubated sections and whole mounts. Intra ... More
Isolation and chemistry of the mixed anhydride intermediate in the reaction catalyzed by dethiobiotin synthetase.
AuthorsGibson KJ
JournalBiochemistry
PubMed ID9214291
'Dethiobiotin synthetase (DTBS) catalyzes the formation of the cyclic urea, dethiobiotin (DTB), from (7R,8S)-diaminononanoic acid (DAPA), CO2, and ATP; the other products of the reaction are ADP and Pi. The first intermediate in the reaction sequence is the 7-carbamate of DAPA [Huang, W., et al. (1995) Biochemistry 34, 10985-10995; Gibson, ... More
Dynamics of trigger factor interaction with translating ribosomes.
AuthorsRutkowska A, Mayer MP, Hoffmann A, Merz F, Zachmann-Brand B, Schaffitzel C, Ban N, Deuerling E, Bukau B,
JournalJ Biol Chem
PubMed ID18045873
'In all organisms ribosome-associated chaperones assist early steps of protein folding. To elucidate the mechanism of their action, we determined the kinetics of individual steps of the ribosome binding/release cycle of bacterial trigger factor (TF), using fluorescently labeled chaperone and ribosome-nascent chain complexes. Both the association and dissociation rates of ... More
Use of a biomimetic peptide in the design of a competitive binding assay for biotin and biotin analogues.
AuthorsGregory KJ, Bachas LG
JournalAnal Biochem
PubMed ID11161298
'A competitive binding assay for biotin, biocytin, and desthiobiotin utilizing a genetically engineered enzyme-ligand conjugate is described herein. This assay is unique in that the enzyme-ligand conjugate consists of the streptavidin binding peptide Strep-tag II, which mimics the binding of biotin to streptavidin, rather than biotin itself. This allows for ... More
Biosynthesis of biotin from dethiobiotin by the biotin auxotroph Lactobacillus plantarum.
AuthorsBowman WC, DeMoll E
JournalJ Bacteriol
PubMed ID8244941
'Lactobacillus plantarum requires biotin for growth. We show that in the presence of high levels of the biotin biosynthetic precursor, dethiobiotin, L. plantarum synthesizes biotin and grows in medium with dethiobiotin but without biotin. Lactobacillus casei also grew under similar conditions.' ... More
Flavodoxin is required for conversion of dethiobiotin to biotin in Escherichia coli.
AuthorsIfuku O, Koga N, Haze S, Kishimoto J, Wachi Y
JournalEur J Biochem
PubMed ID8076639
'We have reported [Ifuku, O., Kishimoto, J., Haze, S., Yanagi, M. & Fukushima, S. (1992) Biosci. Biotechnol. Biochem. 56, 1780-1785] the enzymic conversion of dethiobiotin to biotin (catalyzed by the enzyme encoded by bioB) in cell-free extract of Escherichia coli which had been genetically engineered for high bioB expression. An ... More
COMBINABILITY OF AVIDIN AND STREPTAVIDIN WITH ANALOGS OF BIOTIN.
AuthorsLICHSTEIN HC, BIRNBAUM J
JournalBiochem Biophys Res Commun
PubMed ID14341938
Selection of streptavidin binders from a DNA-encoded chemical library.
AuthorsDumelin CE, Scheuermann J, Melkko S, Neri D
JournalBioconjug Chem
PubMed ID16536467
DNA-encoded libraries of small organic molecules facilitate the construction of large, encoded self-assembling chemical libraries for the identification of high-affinity binders to protein targets. We have constructed a library of 477 chemical compounds, coupled to 48mer-oligonucleotides, each containing a unique six-base sequence serving as "bar-code" for the identification of the ... More
Intermolecular forces and energies between ligands and receptors.
AuthorsMoy VT, Florin EL, Gaub HE
JournalScience
PubMed ID7939660
The recognition mechanisms and dissociation pathways of the avidin-biotin complex and of actin monomers in actin filaments were investigated. The unbinding forces of discrete complexes of avidin or streptavidin with biotin analogs are proportional to the enthalpy change of the complex formation but independent of changes in the free energy. ... More
Highly purified biotin synthase can transform dethiobiotin into biotin in the absence of any other protein, in the presence of photoreduced deazaflavin.
AuthorsMéjean A, Bui BT, Florentin D, Ploux O, Izumi Y, Marquet A
JournalBiochem Biophys Res Commun
PubMed ID8554581
Biotin synthase from Bacillus sphaericus has been purified to homogeneity from a recombinant strain. The UV-visible spectrum of the pure protein reveals the presence of a [2Fe-2S] cluster. The enzyme is active in the conversion of dethiobiotin to biotin in vitro, in the presence of NADPH, AdoMet and additional unidentified ... More
Nanowire-transistor based ultra-sensitive DNA methylation detection.
Accurate detection of DNA methylation at specific gene transcription sites is important to identify potential tumor formation since this epigenetic alteration may result in silencing of tumor suppressor genes that protect against tumor formation or that repair damaged DNA. Current technologies used in DNA methylation detection are complicated and time ... More
Strep-tag II affinity purification: an approach to study intermediates of metalloenzyme biosynthesis.
AuthorsMaier T, Drapal N, Thanbichler M, Böck A
JournalAnal Biochem
PubMed ID9606145
Complex metalloenzymes (e.g., nitrogenase, hydrogenase, urease) are synthesized starting from the apoprotein via several intermediates by the action of accessory proteins. The isolation and biochemical characterization of such intermediates is hampered by their low abundance and their lability. Here we describe a technique for efficient single-step purification of a hydrogenase ... More
Spectroscopic changes during a single turnover of biotin synthase: destruction of a [2Fe-2S] cluster accompanies sulfur insertion.
AuthorsUgulava NB, Sacanell CJ, Jarrett JT
JournalBiochemistry
PubMed ID11444982
Biotin synthase catalyzes the insertion of a sulfur atom between the saturated C6 and C9 carbons of dethiobiotin. Catalysis requires AdoMet and flavodoxin and generates 5'-deoxyadenosine and methionine, suggesting that biotin synthase is an AdoMet-dependent radical enzyme. Biotin synthase (BioB) is aerobically purified as a dimer of 38.4 kDa monomers ... More
The binding of biotin analogues by streptavidin: a Raman spectroscopic study.
AuthorsTorreggiani A, Fini G
JournalBiospectroscopy
PubMed ID9639110
Raman spectra of anhydrous complexes of streptavidin (Strep) with biotin (Bio) and some Bio analogues [Biotin methyl ester (MEBio), desthiobiotin (DEBio), 2'-iminobiotin (IMBio), and diaminobiotin (DABio)] were recorded. The vibrational results indicate that the interaction with some of these ligands is able to modify the overall structure of the protein ... More
Uptake of biotin by human hepatoma cell line, Hep G2: a carrier-mediated process similar to that of normal liver.
AuthorsSaid HM, Ma TY, Kamanna VS
JournalJ Cell Physiol
PubMed ID7525615
Little is known about the cellular and molecular regulation of the uptake process of the water-soluble vitamin biotin into liver cells, the major site of biotin utilization and metabolism. Such studies are best done using a highly viable and homogeneous cellular system that allows examination of prolonged exposure to an ... More
Uptake of biotin by isolated rat liver mitochondria.
This study examined the mechanism of biotin uptake by liver mitochondria. Mitochondria were isolated from rat liver by an established procedure and demonstrated normal respiratory control ratios. Uptake of biotin with time at incubation buffer pH 6.1 was rapid and linear and occurred with a distinct "overshoot" phenomenon that peaked ... More
Syntheses of biotinylated and dethiobiotinylated insulins.
The 600-MHz proton spectrum of dethiobiotin (prepared from d-biotin with Raney nickel) was measured in order to gain information pertaining to its stereochemical homogeneity. The spectrum demonstrated clearly that the material is a 6:1 mixture of two stereoisomers. The cis compound, corresponding to the stereochemistry of d-biotin, is the major ... More